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Molecular and Cellular Biology, December 1999, p. 8559-8569, Vol. 19, No. 12
Department of Biological Chemistry,
University of California, Irvine, Irvine, California
92697-1700,1 and Department of
Anatomy and Cell Biology, University of Florida, Gainesville, Florida
326102
Received 14 July 1999/Returned for modification 30 August
1999/Accepted 7 September 1999
Strains of the yeast Saccharomyces cerevisiae defective
in transcription factor UAF give rise to variants able to grow by transcribing endogenous ribosomal DNA (rDNA) by RNA polymerase II (Pol
II). We have demonstrated that the switch to growth using the Pol II
system consists of two steps: a mutational alteration in UAF and an
expansion of chromosomal rDNA repeats. The first step, a single
mutation in UAF, is sufficient to allow Pol II transcription of rDNA.
In contrast to UAF mutations, mutations in Pol I or other Pol I
transcription factors can not independently lead to Pol II
transcription of rDNA, suggesting a specific role of UAF in preventing
polymerase switch. The second step, expansion of chromosomal rDNA
repeats to levels severalfold higher than the wild type, is required
for efficient cell growth. Mutations in genes that affect recombination
within the rDNA repeats, fob1 and sir2,
decrease and increase, respectively, the frequency of switching to
growth using Pol II, indicating that increased rDNA copy number is a
cause rather than a consequence of the switch. Finally, we show that
the switch to the Pol II system is accompanied by a striking alteration
in the localization and morphology of the nucleolus. The altered state
that uses Pol II for rDNA transcription is semistable and heritable
through mitosis and meiosis. We discuss the significance of these
observations in relation to the plasticity of rDNA tandem repeats and
nucleolar structures as well as evolution of the Pol I machinery.
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Transcription Factor UAF, Expansion and Contraction
of Ribosomal DNA (rDNA) Repeats, and RNA Polymerase Switch in
Transcription of Yeast rDNA
*
Corresponding author. Mailing address: Department of
Biological Chemistry, 240D Med Sci I, University of California, Irvine, Irvine, CA 92697-1700. Phone: (949) 824-4564. Fax: (949)
824-3201. E-mail: mnomura{at}uci.edu.
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